Characterization of glucose transport and glucose transporters in the human choriocarcinoma cell line, BeWo.

Abstract

In this study we have characterized 2-deoxyglucose (2DG) transport and hexose transporter expression in the human choriocarcinoma cell line, BeWo. 2DG uptake in BeWo cells displayed saturable kinetics (V(max), 29+/-1.5 nmol/min/mg protein;K(m), 1.5+/-0.02 m m) and was significantly inhibited in the presence of 2-deoxyglucose, mannose and 3- O -methyl glucose (all at a competing concentration of 30 m m) by up to 97 per cent, but not by galactose or fructose. Glucose uptake was not Na(+)-dependent, but was inhibited by cytochalasin B (by approx 85 per cent) indicating that hexose uptake was mediated via a facilitative glucose transport mechanism. Northern and immunoblot analyses revealed that BeWo cells expressed GLUT1 and GLUT5, but not GLUT2 or GLUT3. On immunoblots, GLUT1 migrated as a broad protein band on SDS-gels (average M(r)of 55 kDa) and treatment with N -glycanase resulted in a significant shift in its electrophoretic mobility; the core protein migrating as a 40 kDa band indicating that the carrier was heavily glycosylated. GLUT5 was detected as a discrete 60 kDa band and like GLUT1, the observed immunoreactive signal was lost when using antiserum that had been pre-adsorbed with the antigenic peptide. Our findings indicate that BeWo cells express a facilitative glucose transport system with characteristics broadly similar to those reported in isolated human placental membrane vesicles and that they are likely to serve as a useful experimental system for studying the regulation of placental glucose transport and transporter expression.

• Authors: S W Shah, H Zhao, S Y Low, H J Mcardle, H S Hundal
• Journal: Placenta (Placenta) Vol. 20 Issue 8 Pg. 651-9 (Nov 1999) ISSN: 0143-4004 [Print] Netherlands
• PMID: 10527819 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 1999 Harcourt Publishers Ltd.

Chemical References

• Glucose Transporter Type 1
• Glucose Transporter Type 5
• Monosaccharide Transport Proteins
• SLC2A1 protein, human
• Deoxyglucose
• Glucose

Topics

• Biological Transport (physiology)
• Choriocarcinoma (metabolism)
• Deoxyglucose (metabolism)
• Female
• Glucose (pharmacokinetics)
• Glucose Transporter Type 1
• Glucose Transporter Type 5
• Humans
• Monosaccharide Transport Proteins (metabolism)
• Placenta (metabolism)
• Pregnancy
• Tumor Cells, Cultured